【摘要】 为了探究干扰Zfx基因对羊冷冻精液品质及精子发生的影响，试验以2只湖羊种公羊为研究对象，通过构建慢病毒干扰载体并采用睾丸注射法(随机挑选1只注射空载体作为对照组，另1只注射浓缩后慢病毒液作为试验组)对解冻后的湖羊精液品质进行检测，采用qRT-PCR方法检测各组精子Zfx、Zfy基因和精子发育相关基因(STRA8、PLCZ1、SYCP3、SPATA19)相对表达量。结果表明:包装后慢病毒滴度为3.3×10~7TU/m L。试验组的单次射精量、精液密度、精子活力均极显著低于对照组(P<0.01)，顶体不完整性、质膜不完整性、精子畸形率均极显著高于对照组(P<0.01)。试验组Zfy、STRA8、PLCZ1基因相对表达量与对照组差异不显著(P>0.05); Zfx、SYCP3、SPATA19基因相对表达量极显著低于对照组(P<0.01)，分别降低了77.07%、22.75%、32.31%。说明Zfx基因在精子发育过程中发挥重要作用，干扰Zfx基因显著影响精子活力及精子发育相关基因表达。更多还原

【Abstract】 In order to explore the effects of Zfx gene interference on the quality of frozen semen and spermatogenesis of sheep,two Hu sheep rams were used as the research objects. The lentivirus interference vector was constructed and the semen quality of the thawed Hu sheep was detected by testicular injection( one was randomly selected to inject empty vector as the control group,and the other was injected with concentrated lentivirus solution as the experimental group). The relative expression levels of Zfx,Zfy gene and sperm development related genes( STRA8,PLCZ1,SYCP3,SPATA19) in each group were detected by qRT-PCR.The results showed that the packaged lentiviral titer was3. 3×10~7 TU/m L. The single ejaculate volume,semen density,and sperm motility were significantly lower than those in the control group( P<0. 01),while acrosome integrity,plasma membrane integrity,and sperm abnormality rate were significantly higher than those in the control group( P<0. 01). There was no significant difference in the relative expression levels of Zfy,STRA8,and PLCZ1 genes( P > 0. 05). The relative expression levels of Zfx,SYCP3,and SPATA19 genes were significantly lower than those in the control group( P<0. 01),decreasing by 77. 07%,22. 75%,and 32. 31%,respectively. The results indicated that the Zfx gene played a crucial role in spermatogenesis,and interfering with the Zfx significantly affected sperm motility and the expression of genes related to sperm development.更多还原